| ObjectiveTo identify the different metabolites between pre-diabetes(Pre-DM) and type2diabetes mellitus(T2DM) with qi-yin deficiency and asymptomatic in traditional chinses medicine based on GC/MS, and find out potential metabolism pathways and signaling pathways in T2DM upon those different metabolites.MethodsSerum samples of35Pre-DM,35T2DM and35healthy volunteers were collected. According to TCM theory,7patients of Pre-DM were qi-yin deficiency and28were asymptomatic,6patients of T2DM were qi-yin deficiency and29were asymptomatic. The samples were detected with Gas Chromatography-tandem mass spectrometry(GC/MS), and principal components analysis(PCA) and PLS—discrimination analysis (PLS-DA) were performed to find out different metabolites between them which were known as potential biomarkers. The potential biomarkers then were analysised by ingenuity pathway analysis (IPA) to study the interaction between them, and potential metabolism pathways and signaling pathways of Pre-DM and T2DM were acquired.Results1. There was no significant difference of sex, uric acid and blood fat between Pre-DM, DM and healthy volunteers, but age, weight, BMI, HbAlc, FPG and2h-PG were significant difference between the three groups.2. PCA scores plots could discriminate T2DM and healthy volunteers significantly, but not well between Pre-DM and health. And there were different metabolites among the three groups.(1) Up regulated potential biomarkers in Pre-DM were:2-Hydroxybutanoic acid, urea, glucose, palmitic acid, galactose, stearic acid, and down regulated biomarker was glycine.(2) Up regulated potential biomarkers in T2DM were:2-Hydroxybutanoic acid,3-Hydroxybutanoic acid, urea, propanetriol, phenylalanine, ornithine, glucose, tyrosine, palmitic acid, galactose, unknown(rt39.7), linoleic acid, oleicacid, stearic acid, and down regulated biomarkers was glycine.(3) During the progression of Pre-DM to T2DM, up regulated metabolites were2-Hydroxybutanoic acid, urea, propanetriol, ornithine, glucose, palmitic acid, galactose, linoleic acid, oleic acid, stearic acid, and down regulated metabolite was:erythrose.(4) The common up regulated metabolites of Pre-DM and T2DM were2-Hydroxybutanoic acid, urea, glucose, palmitic acid, galactose, stearic acid, and the common down regulated metabolites was glycine. Metabolites changes in OGTT2h were similar to free state, but there were more kinds of metabolites in fatty acids and amino acids participate in during the change.3. It could be discriminated from OGTT2h to free states among each of the three groups, up regulated metabolites were glocose and galactose, down regulated metabolites were mainly free fatty acids, cholesterol, amino acids and organic acids.4. The metabolites remained similar changes between qi-yin deficiency and asymptomatic in both Pre-DM and T2DM, GC/MS showed no significant differences between qi-yin deficiency and asymptomatic.5. IPA software was used to analysis the bioinformatics of metabolites. The probable classical metabolism pathways in both Pre-DM and T2DM were cyanoamino acid metabolism, urea cycle and metabolism of amino groups, glutathione metabolism, aminoacyl-tRNA biosynthesis, lysine degradation, glycine serine and threonine metabolism, arginine and proline metabolism, pentose phosphate pathway, galactose metabolism, propanoate metabolism, glycolysis/gluconeogenesis, purine metabolism, starch and sucrose metabolism, pyrimidine metabolism, methane metabolism, nitrogen metabolism, bile acid biosynthesis, fatty acid biosynthesis, fatty acid elongation in mitochondria, fatty acid metabolism. The probable classical metabolism pathways in T2DM were tyrosine metabolism, phenylalanine tyrosine and tryptophan biosynthesis, butanoate metabolism, glycerolipid metabolism, synthesis and degradation of ketone bodies, linoleic acid metabolism. The probable classical metabolism pathways in Pre-DM were biosynthesis of steroids, C21-steroid hormone metabolism.The probable classical signaling pathways in both Pre-DM, T2DM and T2DM vs Pre-DM were growth hormone signaling, type2diabetes mellitus signaling, maturity onset diabetes of young(MODY) signaling, IL-12signaling and production in macrophages, mitochondrial dysfunction, insulin receptor singaling, FXR/RXR activation, glutamate receptor signaling, dopamine-DARPP32feedback in cAMP signaling. The probable classical signaling pathways in T2DM were protein kinase signaling, dopamine receptor signaling. The probable classical signaling pathways in Pre-DM were basal cell carcinoma signaling, role of lipids/lipid rafts in the pathogenesis of influenza, caveolar-mediated endocytosis signaling, virus entry via endocytic pathways, LXR/RXR activation.Conclusions1. Potential metabolites were obtained between Pre-DM and healthy volunteers based on GC/MS, PCA scores plots had the trends to discriminate these two groups.2-Hydroxybutanoic acid, urea, glucose, palmitic acid, galactose and stearic acid were up regulated in Pre-DM, glycine was down regulated.2. potential metabolites were obtained between T2DM and healthy volunteers based on GC/MS, PCA scores plots could discriminate these two groups clearly.2-Hydroxybutanoic acid,3-Hydroxybutanoic acid, urea, propanetriol, phenylalanine, ornithine, glucose, tyrosine, palmitic acid, galactose, unknown (rt39.7), linoleic acid, oleic acid and stearic acid were up regulated in T2DM, glycine was down regulated.3. GC/MS showed no significant differences between qi-yin deficiency and asymptomatic in both Pre-DM and T2DM, and there was no significant difference metabolites of these two subgroups. It indicated that there was similar pathology between them, and in the treatment of asymptomatic Pre-DM or T2DM, tonifying qi and yin could be performed. It provided theory evidence of clinical treatment.4. IPA software was used to analysis the bioinformatics of metabolites. The metabolism pathways and signaling pathways related to discoved metabolites might be key pathways of Pre-DM and T2DM, and it deserved further more studies.
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